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Biomedical subjects

J Wu

Publications and source records attributed to J Wu.

At least 937 records · Page 52Linked to original sources

Follicular dendritic cells and presentation of antigen and costimulatory signals to B cells.

This review focuses on how immunogens trapped by FDC in the form of Ag-Ab complexes productively signal B cells. In vitro. Ag-Ab complexes are poorly immunogenic but in vivo immune complexes elicit potent recall responses. FDC trap Ag-Ab complexes and make immune complex coated bodies or "iccosomes". B cells endocytose iccosomes, the Ag is processed, and T-cell help is elicited. In vitro, addition of FDC bearing appropriate Ag-Ab complex to memory T and B cells provoke potent recall responses (IgG and IgE). FDC also provide nonspecific costimulatory signals which augment B-cell proliferation and Ab production. B cell-FDC contact is important and interference with ICAM-1-LFA-1 interactions reduces FDC-mediated costimulation. Preliminary data suggest that a costimulatory signal may be delivered via CR2L on FDC binding CR2 on B cells. FDC can also stimulate B cells to become chemotactically active and can protect lymphocytes from apoptosis. FDC also appear to be rich in thiol groups and may replace reducing compounds such as 2 mercaptoethanol in cultures. In short, FDC-Ag specifically signals B cells through BCR, and FDC provide B cells with iccosomal-Ag necessary for processing to elicit T-cell help. In addition, FDC provide nonspecific signals that are important to promote B-cell proliferation, maintain viability, and induce chemotactic responsiveness.

Animals↗

Cell death and longevity: implications of Fas-mediated apoptosis in T-cell senescence.

Two prominent features of immune senescence are altered T-cell phenotype and reduced T-cell response. We have previously shown that T-cell senescence is greatly reduced in CD2-fas transgenic mice, in which the Fas apoptosis molecule is constantly expressed on T cells. Using a different experimental approach, the relationship between T-cell senescence and apoptosis was analyzed on human peripheral blood mononuclear cells. The results indicate that there was increased apoptosis of CD45RO- (CD45RA+) T cells upon activation. We propose that this could account for the increase in CD45RO+ "memory" T cells with aging in humans. T-cell responsiveness remained high in CD2-fas transgenic aged mice, but there was no increase in overall life span of these mice. Increased T-cell responsiveness was associated with an increased acute-phase response and serum amyloid A deposition in the glomerulus of aged CD2-fas transgenic mice. Therefore, restoration of the T-cell immune function using a CD2-fas transgene produced undesirable side-effects to aged transgenic mice. In addition to its role in activation-induced cell death, Fas-mediated apoptosis may be important in deleting T cells in response to DNA damage. It may also inhibit cell-cycle progression by cleaving various kinases and DNA repair enzymes. We observed that cell lines derived from human premature aging diseases have a higher sensitivity to Fas-mediated apoptosis. The implications of these observations are discussed.

Acute-Phase Reaction↗

The Caulobacter heat shock sigma factor gene rpoH is positively autoregulated from a sigma32-dependent promoter.

Sigma factor sigma32, encoded by rpoH, is required for the recognition of heat shock genes during normal growth conditions and in response to heat shock and other stresses. Unlike the well-studied Escherichia coli rpoH gene, which is transcribed from four promoters recognized by either a sigma70 (sigmaD)- or sigma24 (sigmaE)-containing RNA polymerase, the Caulobacter crescentus rpoH gene is transcribed from two promoters, P1 and P2. In this study, we have examined the structure and expression of these promoters and shown that the rpoH P2 promoter is sigma32 dependent. We present evidence here that P2 is specifically recognized and transcribed by the reconstituted C. crescentus Esigma32 RNA polymerase holoenzyme. We show that site-directed mutations within either the -10 or the -35 regions of P2 have substantial effects on the levels of transcription by the Esigma32 polymerase predicted from the sigma32 promoter consensus sequence. The mutations have similar effects in vivo as assayed with rpoH-lacZ transcription fusions. Analysis of the rpoH P1 promoter provided evidence that it is sigma70 dependent. S1 nuclease protection assays of rpoH P1- and P2-specific expression after heat shock at 42 or 50 degrees C and during synchronous cell division cycles under normal growth conditions showed that the two promoters are differentially regulated. Mutations within the rpoH P2 promoter consensus sequences abolished the response to heat shock induction in C. crescentus. We conclude from these results that, unlike rpoH genes studied previously in other bacteria, the major transcriptional response of the C. crescentus rpoH gene to heat shock depends on positive autoregulation of the sigma32-dependent promoter.

Bacterial Proteins↗

Aspergillus asexual reproduction and sexual reproduction are differentially affected by transcriptional and translational mechanisms regulating stunted gene expression.

The Stunted protein (StuAp) is a member of a family of transcription factors that regulate fungal development and cell cycle progression. Regulated stuA gene expression is required for correct cell pattern formation during asexual reproduction (conidiation) and for initiation of the sexual reproductive cycle in Aspergillus nidulans. Transcriptional initiation from two different promoters yields overlapping mRNAs (stuA alpha and stuAbeta) that upon translation yield the same protein. Here we show that multiple regulatory mechanisms interact to control (i) developmental competence-dependent expression of both transcripts and (ii) induction-dependent expression of stuA alpha, but not stuAbeta, by the conidiation-specific Bristle (BrlAp) transcriptional activator. Quantitative levels of both mRNAs are further modulated by (i) an activator(s) located at a far-upstream upstream activation sequence, (ii) feedback regulation by StuAp, and (iii) positive translational regulation that requires the peptide product of a micro-open reading frame unique to the stuA alpha mRNA 5' untranslated region. Gradients in stuA alpha expression were most important for correct cell and tissue type development. Threshold requirements were as follows: metula-phialide differentiation < ascosporogenesis < cleistothecial shell-Hülle cell differentiation. Altered stuA expression affected conidiophore morphology and conidial yields quantitatively but did not alter the temporal development of cell types or conidiophore density. By contrast, the sexual cycle showed both temporal delay and quantitative reduction in the number of cleistothecial initials but normal morphogenesis of tissue types.

Amino Acid Sequence↗

Molecular cloning and characterization of human JNKK2, a novel Jun NH2-terminal kinase-specific kinase.

At least three mitogen-activated protein kinase (MAPK) cascades were identified in mammals, each consisting of a well-defined three-kinase module composed of a MAPK, a MAPK kinase (MAPKK), and a MAPKK kinase (MAPKKK). These cascades play key roles in relaying various physiological, environmental, or pathological signals from the environment to the transcriptional machinery in the nucleus. One of these MAPKs, c-Jun N-terminal kinase (JNK), stimulates the transcriptional activity of c-Jun in response to growth factors, proinflammatory cytokines, and certain environmental stresses, such as short wavelength UV light or osmotic shock. The JNKs are directly activated by the MAPKK JNKK1/SEK1/MKK4. However, inactivation of the gene encoding this MAPKK by homologous recombination suggested the existence of at least one more JNK-activating kinase. Recently, the JNK cascade was found to be structurally and functionally conserved in Drosophila, where DJNK is activated by the MAPKK DJNKK (hep). By a database search, we identified an expressed sequence tag (EST) encoding a portion of human MAPKK that is highly related to DJNKK (hep). We used this EST to isolate a full-length cDNA clone encoding a human JNKK2. We show that JNKK2 is a highly specific JNK kinase. Unlike JNKK1, it does not activate the related MAPK, p38. Although the regulation of JNKK1 activities and that of JNKK2 activities could be very similar, the two kinases may play somewhat different regulatory roles in a cell-type-dependent manner.

Amino Acid Sequence↗

Comparative protection against oxyradicals by three flavonoids on cultured endothelial cells.

Oxygen-derived free radicals are known to injure the endothelium of aorta in diverse disorders. In this study we compared the cytoprotective effects of three flavonoids against oxyradical damage to porcine aortic endothelial cells in vitro. Cultured porcine aortic endothelial cells were exposed to oxyradicals generated by xanthine oxidase--hypoxanthine (XO-HP). The cytoprotective activities of morin, quercetin, and catechin on these systems were compared using established morphologic criteria. The results in the XO-HP system showed that morin at 0.125, 0.25, and 0.5 mM delayed cell necrosis to 27.4 +/- 1.3, 46.8 +/- 1.8, and longer than 70 min, respectively, compared with 12.0 +/- 1.3 min in the control group. These degrees of protection were significantly stronger than those provided by quercetin and catechin at corresponding concentrations (p < 0.01). Morin and quercetin were moderate inhibitors of xanthine oxidase on the basis of the oxygen consumption rate, whereas catechin at the same concentrations had little inhibitory effect. The data from uric acid formation and cytochrome c reduction were consistent with the oxygen consumption measurement for the three flavonoids.

Animals↗

Conditions for the induction of long-term potentiation and long-term depression by conjunctive pairing in the dentate gyrus in vitro.

Conditions for the induction of long-term potentiation and long-term depression by conjunctive pairing in the dentate gyrus in vitro. J. Neurophysiol. 78: 2569-2573, 1997. The conditions under which long-term potentiation (LTP) and long-term depression (LTD) of excitatory postsynaptic currents were induced by the conjunctive pairing-type protocol of afferent stimulation and postsynaptic depolarization were studied in the medial perforant pathway-granule cell synapse of the dentate gyrus in vitro. The conjunctive pairing of 1-Hz afferent stimulation and steady state postsynaptic depolarization to 0 mV did not induce LTP or LTD. Inhibition of LTD induction with a phosphatase inhibitor or ruthenium red resulted in induction of LTP after the conjunctive pairing. Such LTP induction was N-methyl--aspartate dependent. Conversely, inhibition of LTP induction with a kinase inhibitor resulted in LTD induction after the conjunctive pairing. Thus the failure to induce LTP or LTD with the pairing protocol involving depolarization to 0 mV membrane potential was due to simultaneous activation of intracellular processes that generate the induction of LTP and LTD. Increasing the frequency of afferent stimulation to 200 Hz, even for just eight stimuli, resulted in LTP induction. The studies show that two factors govern the induction of LTP/LTD, membrane potential and frequency of afferent stimulation, with either increased depolarization or increased afferent stimulation favoring LTP induction.

2-Amino-5-phosphonovalerate↗

Thrombin inhibits atrial natriuretic peptide receptor activity in cultured bovine endothelial cells.

Thrombin and the atrial natriuretic peptide (ANP) possess a number of functionally antagonistic properties in vascular endothelial cells. Thus, regulatory interactions that modulate the activity of one or the other could have important sequelae with regard to cardiovascular homeostasis. Thrombin treatment effected a dose- and time-dependent reduction in ANP receptor activity (maximal 70% to 80% inhibition) in cultured bovine aortic endothelial cells. This resulted from a decrease in total receptor number as well as a modest reduction in the affinity of the receptor for its ligand. The inhibition was largely confined to the type C receptor population, in that thrombin had no effect on maximal type A receptor-linked cGMP accumulation. The protein kinase C-activating phorbol ester 12-O-tetradecanoylphorbol 13-acetate effected a similar reduction in binding activity; however, suppression of protein kinase C activity did not reverse the thrombin effect. Pretreatment of endothelial cells with cycloheximide did not completely prevent the thrombin-dependent inhibition, and thrombin did not effect a reduction in type C receptor mRNA levels, findings that argue for a postsynthetic inhibitory locus. The inhibition of receptor activity was effectively irreversible in that suspension of protein synthesis blocked the recovery of receptor density on the cell surface. Reduction in type C receptor density was accompanied by modest increases in the stability of ANP in the culture medium and enhancement of the cellular cGMP response to the peptide, particularly at low ligand concentrations. These findings demonstrate a potentially important interaction between these two agonist systems in regulating endothelial cell function within the vascular wall.

Amino Acid Chloromethyl Ketones↗

Infiltration of eosinophils into the asthmatic airways caused by interleukin 5.

Interleukin (IL)-5 is thought to play an important role in asthmatic bronchial mucosal inflammation and is a potential therapeutic target. To investigate the effect of IL-5 on the infiltration of eosinophils in airway in vivo, we compared eosinophil counts and their activation status in airways without and after the topical instillation of recombinant human IL-5. Eight subjects with mild atopic asthma underwent initial bronchoscopy during which control bronchoalveolar lavage (BAL) fluid as well as bronchial mucosa were obtained, and at the same time, normal saline and IL-5 were administered to two sublobar segments separately. The second bronchoscopy were carried out and samples from challenged sites were taken 24 h later. It was found that the total eosinophils (BMK-13+ cells) and the activated eosinophils (EG2+ cells) in bronchial mucosa, the eosinophil numbers in BAL fluid, as well as eosinophil cationic protein (ECP) in BAL fluid from saline-challenged segments were not different from those in unchallenged segments. However, a significant eosinophilia was observed in bronchial mucosa and BAL fluid from IL-5-challenged sites. Eosinophil activation, as assessed by secretion of ECP, was also increased significantly in bronchial mucosa and BAL fluid. The results strongly suggested that IL-5 is capable of inducing eosinophil infiltration into the asthmatic airways, as well as the activation of infiltrating eosinophils.

Adult↗

Decreased anterior cingulate gyrus metabolic rate in schizophrenia.

OBJECTIVE: This study examined glucose metabolism in the anterior and posterior cingulate cortex in schizophrenia. METHOD: Fifty unmedicated male schizophrenic patients and 24 normal men were studied with positron emission tomography. RESULTS: Compared with the normal men, the schizophrenic patients had lower relative metabolic rates in the anterior cingulate and higher rates in the posterior cingulate. CONCLUSIONS: The findings suggest hypofunction in the anterior cingulate cortex in schizophrenia.

Adolescent↗

Potentiation of pirarubicin activity in multidrug resistant cells by rifampicin.

The effect of the anti-tuberculosis drug rifampicin on pirarubicin activity was investigated in multidrug-resistant cells overexpressing P-glycoprotein. Rifampicin increased the sensitivity of pirarubicin to anthracycline-resistant mouse leukemic P388 cells and significantly enhanced the cytotoxicity and intracellular accumulation of pirarubicin in resistant cells, but had no effect in parent cells. By contrast, two other rifamycins, rifamycin B and SV, had no effect on pirarubicin accumulation in resistant cells. Rifampicin also enhanced pirarubicin-induced apoptosis and G2/M blockade on the cell cycle in resistant cells. These results show that rifampicin enhances the cytotoxic action of pirarubicin in resistant cells, at least partly via the inhibition of cellular pirarubicin efflux.

Animals↗

Inhibitory effects of dehydrocorydaline isolated from Corydalis Tuber against type I-IV allergic models.

An alkaloidal component, dehydrocorydaline (DHC) isolated from Corydalis Tuber (tuber of Corydalis turtschaninovii forma yanhusuo), has been screened for activity against types I-IV allergic reactions. In a type I allergic models, DHC at a dose of 0.5 mmol/kg, p.o. inhibited 48 h homologous passive cutaneous anaphylaxis (PCA) in rats, which is related to IgE. DHC also exhibited an inhibitory effect on antigen-induced histamine release from peritoneal mast cells. In a type II allergic model, DHC did not inhibit reversed cutaneous anaphylaxis (RCA) in rats. In a type III allergic model, DHC showed weak inhibition on direct passive arthus reaction (DPAR) in rats. Furthermore, in a type IV allergic model, DHC had inhibitory effects on the induction phase and effector phase in picryl chloride-induced contact dermatitis (PC-CD). These results indicated that DHC not only inhibits antibody-mediated allergic reactions but also influences cell-mediated allergia reactions, and the inhibitory effect of Corydalis Tuber on allergic reactions may be partially attributed to DHC.

Alkaloids↗

Photon migration of near-diffusive photons in turbid media: a Lagrangian-based approach.

We show that light transport in a turbid medium can be described by a path integral with a quasi-particle Lagrangian. The most probable trajectory over which photons can be found can be obtained from this Lagrangian. This approach extends the diffusion approximation to the near-diffusive regime, in which photons travel only a few transport mean free paths.

Diffusion↗

Convolution picture of the boundary conditions in photon migration and its implications in time-resolved optical imaging of biological tissues.

To model the effect of a tissue-air boundary on time-resolved optical measurements, a convolution picture is presented based on the photon migration picture. It is demonstrated that the boundary conditions (either index matched or index mismatched) can be formulated as a spatiotemporal convolution of two terms, with the first being identical to the solution in the infinite medium and the second being independent of the original light source. The conditions under which the spatial convolution part in the second term becomes negligible are also determined thus permitting the complete separation of the two terms by use of a temporal Laplace or Fourier transformation. This result is promising, since it, suggests a method of removing the effect of the boundary conditions in the applications of time-resolved optical imaging (in both the time domain and the frequency domain) in biological tissues.

Diagnostic Imaging↗

Effects of zinc gluconate on nephrotoxicity and glutathione metabolism disorder induced by cis-platin in mice.

To examine the effects of zinc gluconate (ZG) on the nephrotoxicity of cis-platin (CDDP), changes in renal function and glutathione metabolism were investigated in mice. Fifty mice were randomly divided into five equal groups: controls, CDDP group and three ZG plus CDDP groups; the dose of ZG was 100 mg, 140 mg and 180 mg/kg respectively. Mice were sacrificed after 48 hours of CDDP (9 mg/kg) treatment; the levels of serum creatinine (Cr) and blood urea nitrogen (BUN) were measured, the glutathione (GSH) and malondialdehyde (MDA) content and the activities of glutathione peroxidase (GHS-PX) and catalase (CAT) in whole blood and renal tissue were assayed. Compared to the control group, there were significant increases in Cr, BUN and MDA, and decreases in the activities of GSH-PX in the CDDP group in whole blood and renal tissue. Preadministration of ZG at the doses of 140 mg/kg and 180 mg/kg significantly increased the concentration of GSH in whole blood, and decreased the levels of Cr and BUN, and the MDA content in whole blood and renal tissue, compared with the CDDP group. We conclude that preadministration of ZG might improve renal function and glutathione metabolism and reduce the nephrotoxicity of cis-platin.

Animals↗

Angiotensinogen gene expression is stimulated by the cAMP-responsive element binding protein in opossum kidney cells.

It has been reported previously that the addition of isoproterenol or forskolin stimulates the expression of the angiotensinogen (ANG) gene in opossum kidney (OK) 27 cells, an OK cell line with a fusion gene containing the 5'-flanking regulatory sequence of the rat ANG gene fused with a human growth hormone (hGH) gene as a reporter, pOGH (ANG N-1498/+18), permanently integrated into their genomes. To investigate whether the effect of isoproterenol or forskolin on the expression of the ANG gene is mediated via the nuclear 43-kD cAMP-responsive element binding protein (CREB), OK 27 cells were transiently transfected with an expression plasmid containing the cDNA for the 43-kD CREB (pRSV/CREB). The level of expression of the pOGH (ANG N-1498/+18) in OK 27 cells was estimated by the amount of immunoreactive hGH secreted into the culture medium. Transfection of pRSV/CREB alone stimulated the expression of pOGH (ANG N-1498/+18). The addition of isoproterenol or forskolin further enhanced the stimulatory effect of pRSV/ CREB on the expression of pOGH (ANG N-1498/+18). The enhancing effect of isoproterenol was inhibited by the presence of propranolol (an inhibitor of beta-adrenoceptors) and (R)-p-adenosine 3'5'-cyclic monophospho-orthioate (Rp)-cAMP (an inhibitor of cAMP-dependent protein kinase A I and II). Transfection of pRSV/CREB had no effect on the expression of thymidine kinase growth hormone in OK 13 cells, an OK cell line with a fusion gene containing the promoter/enhancer DNA sequence of the viral thymidine-kinase gene fused with an hGH gene as a reporter, thymidine kinase growth hormone, permanently integrated into their genomes. These studies demonstrate that isoproterenol stimulates the expression of ANG gene via the cAMP-dependent protein kinase A and probably via the interaction of the 43-kD CREB with the 5'-flanking region of the ANG gene. Our data indicate that the nuclear 43-kD CREB may have a modulatory role on the expression of the ANG gene in OK cells.

Adrenergic beta-Agonists↗

[An immunohistochemical double labelling study on type IV collagenase and basement membrane in human colonic carcinoma].

OBJECTIVE: To investigate the relationship between type IV collagenase and basement membrane (BM) and the relationship between type IV collagenase and p21 expression. METHODS: Immunohistochemical double labelling technique was used to study paraffin embedded sections of 86 cases of colon carcinoma. RESULTS: The positive rate of type IV collagenase was 83.72% in carcinoma cases and 10% in normal colorectal mucosa (P < 0.01). Intact BM was seen in normal colorectal mucosa while BM was disrupted and fragmentary in carcinoma cases. A close positive correlation was also found between the expression of type IV collagenase and p21 in colonic carcinoma (r = 0.974, P < 0.01). CONCLUSIONS: The results suggest that type IV collagenase plays an important role in the destruction of BM and invasion by colonic carcinoma.

Adenocarcinoma↗

Antagonistic effects of extract from leaves of ginkgo biloba on glutamate neurotoxicity.

AIM: To determine whether the extract of leaves of Ginkgo biloba L (EGb) and several active constituents of EGb have protective effects against glutamate (Glu)-induced neuronal damage. METHODS: Microscopy and image analysis of nucleus areas in the arcuate nuclei (AN) of mice were made. The neuronal viability in primary cultures from mouse cerebral cortex was assessed using MTT [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide] staining and the intracellular free calcium concentration ([Ca2+]i) of single neuron was measured using Fura-2. RESULTS: EGb (2.5 mg.L-1) and its constituent ginkgolide B (Gin B, 2 mg.L-1) protected the neuronal viability against Glu-induced injury, and prevented the Glu-induced elevation in [Ca2+]i. EGb (3-10 mg.kg-1) attenuated the decrease of nucleus areas in arcuate nuclei induced by Glu (1 g.kg-1, s.c.). CONCLUSION: EGb and Gin B prevent neurons from Glu neurotoxicity through reduction of the rise in [Ca2+]i.

Animals↗